[用于检测葡萄球菌肠毒素B和C1的化学发光免疫分析方法的建立]

[Establishment of chemiluminescence immunoassay for detecting Staphylococcal enterotoxin B and C1].

作者信息

Chen Li-jie, Liu Zhen-shi, Dong Bang-quan, Yang Kun, Li Qi, Jin Bo-quan

机构信息

Department of Immunology, Fourth Military Medical University, Xi'an 710032, China.

出版信息

Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2006 Sep;22(5):668-9.

PMID:16948922

Abstract

AIM

To establish chemiluminescence immunoassay (CLIA) for detecting staphylococcal enterotoxin B (SEB) and C1 (SEC1) and compare its sensitivity and stability with ELISA.

METHODS

The anti-SEB and SEC1 monoclonal antibodies (mAb) were purified by Q Sepharose Fast Flow chromatographic column. The alkaline phosphatase (AP) conjugated mAbs FMMU-SEB.D6 and FMMU-SEC1.C4 were used as detecting antibodies and the FMMU-SEB.B4 and FMMU-SEC1.G8 mAbs were used as coating antibodies in both methods. Phenolphthalein monophosphate (PMP) and lumigen APS-5 were employed as substrates for AP in ELISA and CLIA, respectively. The light was detected and measured by the GENios analyzer (TECAN Group Ltd.). The sensitivities and detect ranges of CLIA and ELISA methods were compared.

RESULTS

Compared with ELISA, CLIA was more sensitive (0.1 ng/mL vs 0.39 ng/mL) and timesaving. Furthermore, the liner range of CLIA was broader than that of ELISA (0.78-50 ng/mL vs 3.125-50 ng/mL).

CONCLUSION

CLIA for detecting SEB and SEC1 are established successfully which may be useful in food monitoring, epidemiology survey and detecting SE contaminated samples in environment.

摘要

目的

建立检测葡萄球菌肠毒素B（SEB）和C1（SEC1）的化学发光免疫分析法（CLIA），并将其灵敏度和稳定性与酶联免疫吸附测定（ELISA）法进行比较。

方法

采用Q Sepharose Fast Flow层析柱纯化抗SEB和SEC1单克隆抗体（mAb）。两种方法均使用碱性磷酸酶（AP）偶联的单克隆抗体FMMU-SEB.D6和FMMU-SEC1.C4作为检测抗体，FMMU-SEB.B4和FMMU-SEC1.G8单克隆抗体作为包被抗体。分别以单磷酸酚酞（PMP）和鲁米诺APS-5作为ELISA法和CLIA法中AP的底物。通过GENios分析仪（帝肯集团有限公司）检测和测量发光强度。比较CLIA法和ELISA法的灵敏度和检测范围。